In utero delivery of targeted ionizable lipid nanoparticles facilitates in vivo gene editing of hematopoietic stem cells.

Autor:	Palanki R; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104.; Center for Fetal Research, Division of General, Thoracic, and Fetal Surgery, Children's Hospital of Philadelphia, Philadelphia, PA 19104., Riley JS; Center for Fetal Research, Division of General, Thoracic, and Fetal Surgery, Children's Hospital of Philadelphia, Philadelphia, PA 19104., Bose SK; Center for Fetal Research, Division of General, Thoracic, and Fetal Surgery, Children's Hospital of Philadelphia, Philadelphia, PA 19104., Luks V; Center for Fetal Research, Division of General, Thoracic, and Fetal Surgery, Children's Hospital of Philadelphia, Philadelphia, PA 19104., Dave A; Center for Fetal Research, Division of General, Thoracic, and Fetal Surgery, Children's Hospital of Philadelphia, Philadelphia, PA 19104., Kus N; Center for Fetal Research, Division of General, Thoracic, and Fetal Surgery, Children's Hospital of Philadelphia, Philadelphia, PA 19104., White BM; Center for Fetal Research, Division of General, Thoracic, and Fetal Surgery, Children's Hospital of Philadelphia, Philadelphia, PA 19104., Ricciardi AS; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104.; Center for Fetal Research, Division of General, Thoracic, and Fetal Surgery, Children's Hospital of Philadelphia, Philadelphia, PA 19104., Swingle KL; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104., Xue L; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104., Sung D; Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104., Thatte AS; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104., Safford HC; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104., Chaluvadi VS; Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104., Carpenter M; Center for Fetal Research, Division of General, Thoracic, and Fetal Surgery, Children's Hospital of Philadelphia, Philadelphia, PA 19104., Han EL; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104., Maganti R; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104.; Center for Fetal Research, Division of General, Thoracic, and Fetal Surgery, Children's Hospital of Philadelphia, Philadelphia, PA 19104., Hamilton AG; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104., Mrksich K; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104., Billingsley MB; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104., Zoltick PW; Center for Fetal Research, Division of General, Thoracic, and Fetal Surgery, Children's Hospital of Philadelphia, Philadelphia, PA 19104., Alameh MG; Department of Pathology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104., Weissman D; Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104., Mitchell MJ; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA 19104., Peranteau WH; Center for Fetal Research, Division of General, Thoracic, and Fetal Surgery, Children's Hospital of Philadelphia, Philadelphia, PA 19104.
Jazyk:	angličtina
Zdroj:	Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2024 Aug 06; Vol. 121 (32), pp. e2400783121. Date of Electronic Publication: 2024 Jul 30.
DOI:	10.1073/pnas.2400783121
Abstrakt:	Monogenic blood diseases are among the most common genetic disorders worldwide. These diseases result in significant pediatric and adult morbidity, and some can result in death prior to birth. Novel ex vivo hematopoietic stem cell (HSC) gene editing therapies hold tremendous promise to alter the therapeutic landscape but are not without potential limitations. In vivo gene editing therapies offer a potentially safer and more accessible treatment for these diseases but are hindered by a lack of delivery vectors targeting HSCs, which reside in the difficult-to-access bone marrow niche. Here, we propose that this biological barrier can be overcome by taking advantage of HSC residence in the easily accessible liver during fetal development. To facilitate the delivery of gene editing cargo to fetal HSCs, we developed an ionizable lipid nanoparticle (LNP) platform targeting the CD45 receptor on the surface of HSCs. After validating that targeted LNPs improved messenger ribonucleic acid (mRNA) delivery to hematopoietic lineage cells via a CD45-specific mechanism in vitro, we demonstrated that this platform mediated safe, potent, and long-term gene modulation of HSCs in vivo in multiple mouse models. We further optimized this LNP platform in vitro to encapsulate and deliver CRISPR-based nucleic acid cargos. Finally, we showed that optimized and targeted LNPs enhanced gene editing at a proof-of-concept locus in fetal HSCs after a single in utero intravenous injection. By targeting HSCs in vivo during fetal development, our Systematically optimized Targeted Editing Machinery (STEM) LNPs may provide a translatable strategy to treat monogenic blood diseases before birth. Competing Interests: Competing interests statement:R.P., M.J.M., and W.H.P. are inventors on a U.S. Provisional Patent Application (No. 63/623,674) related to the technology described in the manuscript.
Databáze:	MEDLINE
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